To study dynamic remodelling of protein complexes, C2C12 myotubes were treated with 0.5 mM palmitic acid for 24 h to induce insulin resistance (IR) and then stimulated with or without acute 100 nM insulin for 30 min followed by PCP-MS(n=3 each. Samples were lysed in non-denaturing buffer, separated by BN-PAGE, and each lane cut into 30 x 1.5mm fractions. Each fraction underwent in-gel digestion and was analysed with liquid chromatography – tandem mass spectrometry (LC-MS/MS) using data-independent acquisition (DIA) resulting in the analysis of 360 individual runs.